Introduction. There is ample evidence that disseminated tumor cells (DTC), which are found in the bone marrow (BM) of patients with breast cancer (BC), including early stages, are progenitors of subsequent distant metastasis. Therefore, BM-DTC represent an additional tool for understanding carcinogenesis and estimating prognosis. Nevertheless, the existing data are controversial. The purpose of the study – to determine the frequency of DTC detection in BM of patients with luminal BC and also its relationship with some clinical and immunophenotypic parameters. Materials and methods. BM bioptates of 65 luminal BC patients were analyzed for the presence of DTC by Attune Acoustic Focusing Cytometer. For the first time in Russia, the sensitivity of the DTC detection method in the BM to the level of 1 × 10–7 myelocaryocytes was increased. Results. In BM, DTC were detected in 40 % of patients, and this finding did not correlate with stage of BC and degree of malignancy. The level of CD8+ lymphocytes in patients with DTC in the BM was significantly lower and amounted to 39,2 % versus 48,1 % in patients without DTC (p = 0,011). The content of myelocaryocytes with DTC-positive status was 1,6 times lower than in the absence of DTC (р = 0,007). Other parameters of the myelogram did not differ significantly. Moreover, no significant correlations were found between the presence of DTC in BM and the breast tumor immunophenotype (HLA-I: p = 0,74; HLA-DR: p = 0,93; CD71: p = 0,46). Conclusion. The presence of BM-DTC is more interrelated with myelogram and subpopulation of BM lymphocytes than with the clinical characteristics of tumor.
Background & Aims. Evaluation of the minimal residual disease (MRD) at different stages of chemotherapy is one of key prognostic factors and a factor of stratification of patients into risk groups in acute lymphoblastic leukemia (ALL). The MRD detection on Day 15 and at later stages is based on identifying blast cells with a leukemia-associated immune phenotype. The aim is to assess the potential of 8-color standardized EuroFlow panels and to detect individual criteria for MRD monitoring during primary diagnosis. Materials & Methods. The analysis included data on the primary immune phenotype and MRD assessment during chemotherapy in 10 adults and 35 children with a confirmed diagnosis of B-cell precursors ALL. Results. The ALL phenotype characteristics at the stage of primary diagnosis permit to make the most complete description of the of 8-color standardized EuroFlow panels. This gives an opportunity to select the most informative antigen combinations for further MRD monitoring. Combinations with CD58/CD38, CD81/CD9 antigen expression, as well as assessment of pan-myeloid CD13, CD33 antigen co-expression may be recommended as the most frequent aberrant immune phenotypes of blast cells in ALL. As for B-lineage progenitor cells in children on Day 15 of the induction therapy, a detection of TdT+ сyCD22<sup>+</sup> cell population is necessary in addition to the quantification of CD10+ and/or CD34<sup>+</sup> В-lineage progenitor cells. Conclusion. Therefore, the 8-color standardized EuroFlow panels permit not only to characterize the primary ALL immune phenotype in details, but may also be widely used for MRD evaluation at all stages of chemotherapy.
Background. Flow cytometry (FC) algorithms of detection of minimal residual disease (MRD) are well standardized, and approximate to molecular biologic methods. However, besides informative leukemia-associated aberrant immunophenotype, which are selected taking into account a tumor phenotype at diagnostics stage, it is necessary to consider specificity of the provided taget therapy and its influence on a cell. Objective: to offer stable combinations of antigens to identify B-cell precursors in patients on therapy of blinatumomab. Materials and methods. Clinical observation of patient G. 4 years old with B-cell precursors acute lymphoblastic leukemia (ALL) (pre-pre-B immunosubtype), whom after 3 bloks of reinduction therapy, taking into account MRD-positive status, blinatumomab was appointed as a monotherapy. Tumor immunophenotype was characterized in details by FC protocol according to EuroFlow in debute and relapse of the disease. MRD monitoring was provided by 8-color FC taking into account personalized leukemia-associated aberrant immunophenotypes. Results. In patient with B-cell precursors ALL received blinatumomab, the strategy of MRD monitoring was changed. Due to the lack of CD19 expression, identification of B-cell precursors was based on expression of cyCD22 in combination with nuclear TdT and CD10. Conclusion. In case of blinatumomab’s appointment during B-cell precursors ALL therapy, it is necessary to change the strategy of B-cell precursors identification, due to the lack of CD19 expression. Detection of B-cell precursors should be provided by assessment of other pan-B lineage antigens. First of all, it is cyCD22 or cyCD79a in combination with nuclear TdT and CD10, within the limits of nucleated cells of the sample.
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